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PYE Medium Recipe

Objective

Author(s): Jessie Luo, Ada Jiang

Prepare Peptone Yeast Extract (PYE) nutrient medium selectively culture Caulobacter crescentus. This protocol describes the preparation of 500 - 1000 mL of medium, adapted from both DSMZ 1462: CAULOBACTER PYE MEDIUM and a protocol provided by our advisor, Beth.

Procedure

Follow the same procedure as LB Medium Recipe, using the following reagents instead:

Version 1 (from standard)
ComponentAmount (for 500 mL)Final concentration
Peptone1 g2 g/L
Yeast extract0.5 g1 g/L
CaCa₂·2H₂O (Calcium chloride dihydrate)0.05 g0.1 g/L
MgSO₄·2H₂O (Magnesium sulfate heptahydrate)0.1 g0.2 g/L
Optional: Agar (for solid medium)6 g1.2%
Version 2 (from manuscript)

0.3% peptone, 0.2% yeast extract, 1.5 mM MgSO4MgSO_4, 1 mM CaCl2CaCl_2, 1.2% agar for solid medium

ComponentAmount (for 1000 mL)Final concentration
Peptone3.00 g3 g/L
Yeast extract2.00 g2 g/L
CaCa₂·2H₂O (Calcium chloride dihydrate)219.08 mg0.219 g/L or 1 mM
MgSO₄·2H₂O (Magnesium sulfate heptahydrate)369.71 mg0.370 g/L or 1.5 mM
Optional: Agar (for solid medium)12.00 g1.2%

Note that the concentration of agar can be increase to 1.5% for a denser gel.